Aptamer Validation by Western Blot–an overview

Western blot is the main and basic technique in cellular and molecular biology. The principle of the western blot is the isolation and detection of the target molecule usually from a cellular extract. The whole process of western blot consists of three stages and can be described briefly as separation of   protein. followed by transportation to a solid membrane and finally detection of the target by an antibody. Western blot technique is usually used for the detection of proteins but also can be used to detect other molecules such as aptamers. Aptamers can be defined as a short-stranded DNA or RNA that bind with the target with high specificity and affinity. Aptamers highly resemble antibodies with many advantages. In this review, there is a focus on the aptamers that had validated by western blot technique other than other methods. This method has the advantage of   less time required, no antibodies needed, and introducing the possibility of multiplexing detection

SIRT1 activators as novel therapy for cancer

Sirutins 1-7 (SIRT1-7) is an enzyme that depends on NAD+ to be activated, making it a member of the 3rd class of Deacetylase enzymes. SIRT1-7’s activity is involved with metabolism, cell survival and/or death as well as DNA repair, gene repression, inflammatory responses, the   aging process, neuroprotection in addition to possibly helping with the treatment of cancer. Molecules that could have a modifying effect on SIRT1-7’s activity has caught a great attention recently, owing to the fact of how beneficial this enzyme could be. In this review, we attempt to shed a light on these activator compounds and their use in Sirutin activation therapy, particularly SIRT1, for it is the most researched type. One of these compounds is Resveratrol, a natural compound that –due to its SIRT 1 activation potential – could help in the treatment of obesity, prevention of tumor formation as well as decrease in heart function and neuronal loss related to aging; however, Resveratrol has poor bioavailability, which is why structurally reformulated compounds and molecules have been developed. Other molecules that are different from Resveratrol such as SRT1720, SRT2104 and SRT2379 in addition to others, have been used and shown greater activation potential for SIRT1 than Resveratrol

Preventive treatment of coronavirus disease-2019 virus using coronavirus disease-2019-receptor-binding domain 1C aptamer by suppress the expression of angiotensin-converting enzyme 2 receptor

The cause of the worldwide coronavirus disease-2019 (COVID-19) pandemic is the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). It is known to employ the same entry portal as SARS-CoV, which is the type 1 transmembrane angiotensin-converting enzyme 2 (ACE2) receptor. The receptor-binding domain (RBD) is located on the spike S-protein's S1 subunit of the spike glycoprotein. The most important and effective therapy method is inhibiting the interaction between the ACE2 receptor and the S-spike RBD. An aptamer is a small, single-chain oligonucleotide that binds strongly to the target molecule. Recently, a CoV-2-RBD-1C aptamer-based system with a 51-base hairpin structure was discovered to have substantial binding affinity against the SARS-CoV-2RBD with similar binding sites at ACE. In the current study, we will study the aptamer's effect as a SARS-CoV-2 spike blocker and inhibit its ACE2 receptors' binding by studying the toxicity of aptamer for this cell line by calcein assay and the inhibition test of CoV-2-RBD-1C aptamers on spike RBD–ACE2 binding. The results show the half-maximum inhibitory concentration of CoV-2-RBD-1C aptamer is 0.08188 μM. The inhibition effect of CoV-2-RBD-1C aptamer on spike RBD–ACE2 binding was determined at half-maximal effective concentration of 0.5 μM concentration. The percentage of spike-ACE2 binding inhibition in A549-hACE2 cells in the D614G variant after 30 s was 77%. This percentage is higher than D614 and N501Y and equals 55% and 65%, respectively, at 0.15 μM of CoV-2-RBD-1C aptamer. The CoV-2-RBD-1C aptamer prevents virus entrance through spike inhibition, which results in a 90% reduction in spike D614 virus transduction at 1.28 μM. In conclusion, the CoV-2-RBD-1C aptamer might be an effective treatment against COVID-19 infection because it directly affects the virus by blocking the S-spike of SARS-CoV-2 and preventing ACE2 receptor binding